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Plasmid Files

pGFP-C-shLenti

HuSH-29 lentiviral vector with chloramphenicol and puromycin resistance markers and a TurboGFP gene, for expressing an shRNA from the U6 promoter.

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pGFP-C-shLenti Sequence and MappGFP-C-shLenti.dna
Map and Sequence File   
Sequence Author:  OriGene
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 AclI (8255) BssS α I (7346) DrdI (7281) Bsu36I (6597) BsrGI (5757) PmeI (5738) BsaBI * (5499) AarI (5487) BlpI (5210) PaeR7I - XhoI (5030) NotI (5024) MluI (5015) AscI (4978) MreI - SgrAI (4974) AsiSI (4972) CMV enhancer SgrDI (1) CMV enhancer EcoNI (1550) AleI (1957) KflI (2314) EcoRI (2618) BamHI (2951) shRNA site BsmBI (3003) SphI (3087) NsiI (3089) SfiI (3220) AvrII (3267) PflFI - Tth111I (3338) BstEII (3430) SexAI * (3863) XbaI (4271) pGFP-C-shLenti 8691 bp
AclI  (8255)
1 site
A A C G T T T T G C A A
BssSαI  (7346)
1 site
C A C G A G G T G C T C
DrdI  (7281)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
Bsu36I  (6597)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BsrGI  (5757)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
PmeI  (5738)
1 site
G T T T A A A C C A A A T T T G
BsaBI  (5499)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AarI  (5487)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BlpI  (5210)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PaeR7I  (5030)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (5030)
1 site
C T C G A G G A G C T C
NotI  (5024)
1 site
G C G G C C G C C G C C G G C G
MluI  (5015)
1 site
A C G C G T T G C G C A
AscI  (4978)
1 site
G G C G C G C C C C G C G C G G
MreI  (4974)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (4974)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
AsiSI  (4972)
1 site
G C G A T C G C C G C T A G C G
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
EcoNI  (1550)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
AleI  (1957)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (2314)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
EcoRI  (2618)
1 site
G A A T T C C T T A A G
BamHI  (2951)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
BsmBI  (3003)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
SphI  (3087)
1 site
G C A T G C C G T A C G
NsiI  (3089)
1 site
A T G C A T T A C G T A
SfiI  (3220)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (3267)
1 site
C C T A G G G G A T C C
PflFI  (3338)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3338)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BstEII  (3430)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
SexAI  (3863)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
XbaI  (4271)
1 site
T C T A G A A G A T C T
TurboGFP
5036 .. 5731  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
5036 .. 5731  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
CmR
7899 .. 8558  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
7899 .. 8558  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
PuroR
3296 .. 3895  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3296 .. 3895  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5875 .. 6463  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5875 .. 6463  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7234 .. 7822  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7234 .. 7822  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4301 .. 4680  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4301 .. 4680  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
CMV promoter
4681 .. 4884  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
4681 .. 4884  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 promoter
3086 .. 3282  =  197 bp
SV40 early promoter
SV40 promoter
3086 .. 3282  =  197 bp
SV40 early promoter
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
6992 .. 7172  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
6992 .. 7172  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
8559 .. 8663  =  105 bp
AmpR promoter
8559 .. 8663  =  105 bp
shRNA site
2951 .. 3008  =  58 bp
shRNA insertion site
shRNA site
2951 .. 3008  =  58 bp
shRNA insertion site
loxP
5786 .. 5819  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
5786 .. 5819  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
T7 promoter
4910 .. 4928  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4910 .. 4928  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
3133 .. 3268  =  136 bp
SV40 origin of replication
SV40 ori
3133 .. 3268  =  136 bp
SV40 origin of replication
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